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乳腺癌和非恶性乳腺组织中MUC5B脱辅基粘蛋白表达的免疫组织化学分析。

Immunohistochemical analysis of MUC5B apomucin expression in breast cancer and non-malignant breast tissues.

作者信息

Sóñora Cecilia, Mazal Daniel, Berois Nora, Buisine Marie-Pierre, Ubillos Luis, Varangot Mario, Barrios Enrique, Carzoglio Julio, Aubert Jean-Pierre, Osinaga Eduardo

机构信息

Departamento de Bioquímica, Laboratorio de Oncología Básica, Universidad de la República, Montevideo, Uruguay.

出版信息

J Histochem Cytochem. 2006 Mar;54(3):289-99. doi: 10.1369/jhc.5A6763.2005. Epub 2005 Sep 7.

Abstract

A deregulation of several MUC genes (MUC1, MUC2, MUC3, MUC5AC, and MUC6) was previously demonstrated in breast carcinomas. Considering that recently we found the "non-mammary" MUC5B mRNA in primary breast tumors (Berois et al. 2003), we undertook the present study to evaluate the expression profile of MUC5B protein product in breast tissues, using LUM5B-2 antisera raised against sequences within the non-glycosylated regions of this apomucin. Expression of MUC5B by breast cancer cells was confirmed by immunocytochemistry, in situ hybridization, and Western blot on MCF-7 cancer cells. Using an immunohistochemical procedure, MUC5B apomucin was detected in 34/42 (81%) primary breast tumors, in 13/14 (92.8%) samples of non-malignant breast diseases, in 8/19 (42.1%) samples of normal-appearing breast epithelia adjacent to cancer, and in 0/5 normal control breast samples. The staining pattern of MUC5B was very different when comparing breast cancer cells (cytoplasmic) and non-malignant breast cells (predominantly apical and in the secretory material). We analyzed MUC5B mRNA expression using RT-PCR in bone marrow aspirates from 22/42 patients with breast cancer to compare with MUC5B protein expression in the primary tumors. Good correlation was observed because the six MUC5B-positive bone marrow samples also displayed MUC5B expression in the tumor. Our results show, for the first time at the protein level, that MUC5B apomucin is upregulated in breast cancer. Its characterization could provide new insights about the glycobiology of breast cancer cells.

摘要

先前已证实在乳腺癌中几种粘蛋白基因(MUC1、MUC2、MUC3、MUC5AC和MUC6)存在失调。鉴于我们最近在原发性乳腺肿瘤中发现了“非乳腺”MUC5B mRNA(Berois等人,2003年),我们开展了本研究,使用针对这种脱辅基粘蛋白非糖基化区域序列产生的LUM5B - 2抗血清来评估乳腺组织中MUC5B蛋白产物的表达谱。通过免疫细胞化学、原位杂交和对MCF - 7癌细胞进行蛋白质印迹法证实了乳腺癌细胞中MUC5B的表达。采用免疫组织化学方法,在42例原发性乳腺肿瘤中的34例(81%)、14例非恶性乳腺疾病样本中的13例(92.8%)、与癌相邻的外观正常的乳腺上皮样本中的19例中的8例(42.1%)以及5例正常对照乳腺样本中的0例中检测到了MUC5B脱辅基粘蛋白。比较乳腺癌细胞(细胞质)和非恶性乳腺细胞(主要在顶端和分泌物质中)时,MUC5B的染色模式非常不同。我们使用逆转录聚合酶链反应(RT - PCR)分析了42例乳腺癌患者中22例的骨髓抽吸物中的MUC5B mRNA表达,以与原发性肿瘤中的MUC5B蛋白表达进行比较。观察到良好的相关性,因为六个MUC5B阳性骨髓样本在肿瘤中也显示出MUC5B表达。我们的结果首次在蛋白质水平表明,MUC5B脱辅基粘蛋白在乳腺癌中上调。其特性可能为乳腺癌细胞的糖生物学提供新的见解。

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